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Proc Natl Acad Sci U S A. 2016 Nov 1;113(44):E6877-E6886. Epub 2016 Oct 17.

Glutamate-induced RNA localization and translation in neurons.

Author information

1
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461.
2
Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461.
3
Gruss Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY 10461.
4
Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA 20147.
5
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461; robert.singer@einstein.yu.edu.

Abstract

Localization of mRNA is required for protein synthesis to occur within discrete intracellular compartments. Neurons represent an ideal system for studying the precision of mRNA trafficking because of their polarized structure and the need for synapse-specific targeting. To investigate this targeting, we derived a quantitative and analytical approach. Dendritic spines were stimulated by glutamate uncaging at a diffraction-limited spot, and the localization of single β-actin mRNAs was measured in space and time. Localization required NMDA receptor activity, a dynamic actin cytoskeleton, and the transacting RNA-binding protein, Zipcode-binding protein 1 (ZBP1). The ability of the mRNA to direct newly synthesized proteins to the site of localization was evaluated using a Halo-actin reporter so that RNA and protein were detected simultaneously. Newly synthesized Halo-actin was enriched at the site of stimulation, required NMDA receptor activity, and localized preferentially at the periphery of spines. This work demonstrates that synaptic activity can induce mRNA localization and local translation of β-actin where the new actin participates in stabilizing the expanding synapse in dendritic spines.

KEYWORDS:

HaloTag; RNA localization; glutamate uncaging; single molecule; β-actin

PMID:
27791158
PMCID:
PMC5098659
DOI:
10.1073/pnas.1614267113
[Indexed for MEDLINE]
Free PMC Article

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